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Anti-NCX2 (SLC8A2) Antibody

Sodium/calcium exchanger 2, Na+/Ca2+ exchange protein 2
Cat #: ANX-012
Alternative Name Sodium/calcium exchanger 2, Na+/Ca2+ exchange protein 2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)RVGDAQGMFEPDGG, corresponding to amino acid residues 486-499 of rat NCX2 (Accession P48768). 3rd intracellular loop.
Accession (Uniprot) Number P48768
Gene ID 140447
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Human, mouse - identical.
RRID AB_2341022.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ic*, ifc*, ip*
Western blot
  • Western blot analysis of rat brain membranes (lanes 1 and 5), mouse brain membranes (lanes 2 and 6), rat skeletal muscle lysate (lanes 3 and 7) and human SH-SY5Y neuroblastoma cell lysate (lanes 4 and 8):
    Western blot analysis of rat brain membranes (lanes 1 and 5), mouse brain membranes (lanes 2 and 6), rat skeletal muscle lysate (lanes 3 and 7) and human SH-SY5Y neuroblastoma cell lysate (lanes 4 and 8):
    1-4. Anti-NCX2 (SLC8A2) Antibody (#ANX-012), (1:200).
    5-8. Anti-NCX2 (SLC8A2) Antibody, preincubated with NCX2/SLC8A2 Blocking Peptide (#BLP-NX012).
Immunohistochemistry
  • Expression of NCX2 in rat cerebellum
    Expression of NCX2 in rat cerebellum
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NCX2 (SLC8A2) Antibody (#ANX-012), (1:100). A. NCX2 staining (red) is expressed in the molecular layer (M) and Purkinje cells (arrows). B. DAPI counterstain (blue) displays the layout of cerebellar layers. C. Merge of A and B demonstrates restriction of NCX2 expression to the molecular layer and Purkinje cells.
References
  1. Lytton, J. (2007) Biochem. J. 406, 365.
  2. Lee, S.H. et al. (2002) J. Neurosci. 22, 6891.
  3. Wanaverbecq, N. et al. (2003) J. Physiol. 550, 83.
  4. Schwarz, E. and Benzer, S. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 10249.
  5. Cai, X. and Lytton, J. (2004) Mol. Biol. Evol. 21, 1692.
  6. Kimura, J. et al. (2009) Biol. Pharm. Bull. 32, 325.
  7. Annunziato, L. et al. (2004) Pharmacol. Rev. 56, 633.
  8. Papa, M. et al. (2003) J. Comp. Neurol. 461, 31.
Scientific background

Ca2+ has proven to be a universal signaling molecule in excitable and non-excitable cells. However, being that its intracellular concentration is 1000 times lower than that of the extracellular milieu, it is important for the cell to keep this ratio for proper function. NCX, a Na+/Ca2+ exchanger is responsible for most of the efflux of Ca2+ from the cell1-3. The NCX transporter is a member of the SLC8 family of solute carriers which in turn belong to the CaCA superfamily1,4,5.

NCX-1 is one of three Na+/Ca2+ exchangers (NCX-1, NCX-2, NCX-3) leading to one Ca2+ movement across the plasma membrane in exchange of three Na+ influx. However, the transporter can reverse the direction of the transport if the concentrations of Na+ and Ca2+ change6. The transporter has nine transmembrane domains and intracellular N- and C-terminals. Between tansmembrane domains 5 and 6, the presence of an extra-long intracellular loop, termed the f-loop, is responsible for regulating the activity of NCX-1 via several different mechanisms like ion binding, phosphorylation, etc. The f-loop also has sites which undergo alternative splicing7.

Of the three NCX expressed in mammalian cells, NCX-1 is the most widely expressed. Its expression is detected in the heart, brain, and kidney. NCX-1 undergoes alternative splicing in a tissue-dependent manner. The first splice region does not change the overall structure of the protein but rather enables the expression of the gene specific to the tissues which require the expression of the gene. The second splicing site leads to a number of proteins varying in length. NCX-2 expression is much more limited; it is expressed only in neurons. NCX-3 is expressed in skeletal muscle and in some regions of the brain and undergoes alternative splicing in a similar fashion to that of NCX-11,8.

Due to their central role in modulating Ca2+ levels in the cell, NCX exchangers are involved in various pathophysiological diseases/disorders such as hypoxia, aging, Alzheimer’s disease7, to name a few.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 16/08/2020

Anti-NCX2 (SLC8A2) Antibody (#ANX-012) is a highly specific antibody directed against an epitope of the rat Na+/Ca2+ exchange protein 2. The antibody can be used in western blot ans immunohistochemistry applications. It has been designed to recognize NCX2 from human, rat, and mouse samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Human glioblastoma LN-229, T98G and U373 cell lysates.
    Song, M. et al. (2014) Br. J. Pharmacol. 171, 3432.
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